Despite extremely effective combination antiretroviral therapy (cART), HIV-1 persists in a small pool of latently infected, resting memory CD4+ T cells. Rapid viral rebound occurs after treatment interruptions. Therefore, life-long antiretroviral therapy is required, raising concerns about adverse effects over decades of therapy, the evolution of drug resistance, and the financial burden of chronic treatment. Current approaches to purging the latent reservoir involve pharmacologic reactivation of HIV-1 transcription by agents that reverse viral latency. The next step is to eliminate infected cells in which HIV-1 gene transcription has been induced by latency reversal agents (LRAs). To date, no broadly applicable strategy has been developed to effectively clear latent HIV-1 in vivo. Two major obstacles have prevented us from developing effective strategies to eliminate latent HIV-1 are: 1) the lack of understanding of cellular composition and anatomical location of the latent viral reservoirs in patients; and 2) the paucity of in vivo experimental systems with which to assess LRA efficacy and vaccination strategies to clear latent HIV-1. I studied HIV-1 infection in recently developed humanized mouse model named MISTRG, which supports co-development of human lymphoid and myeloid lineage cells. I have demonstrated that this novel mouse model could enable us to study HIV-1 infection in CD4+ T cells and multiple subsets of monocytes and macrophages. In the proposed work, I will utilize further improved MISTRG mouse models to systemically investigate HIV-1 latent infection in vivo and develop therapeutic strategies to purge latent viral reservoir. In K99 phase, I will first establish a humanized mouse model of HIV-1 latency under effective cART. With this animal model, I will investigate HIV-1 latent infection i macrophages as well as CD4+ T cells in secondary lymphoid organs and peripheral tissues including gut. Having developed MISTRG model for HIV-1 latent infection, I will study HIV-1 CTL escape variants in the latent reservoir, their response to LRA treatment, and design novel vaccination strategies to boost HIV-1-specific CTL responses at mucosal tissues for viral clearance (R00 phase). Overall, these studies will provide a scientific basis and preclinical evidence of a cure for HIV/AIDS.